CDDO-Me 阻断 Stat3 抑制脊索瘤肿瘤细胞生长。

Blockage of Stat3 with CDDO-Me inhibits tumor cell growth in chordoma.

机构信息

Department of Orthopaedic Surgery, Center for Sarcoma and Connective Tissue Oncology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA.

出版信息

Spine (Phila Pa 1976). 2010 Aug 15;35(18):1668-75. doi: 10.1097/BRS.0b013e3181c2d2b4.

DOI:10.1097/BRS.0b013e3181c2d2b4

PMID:20386502

Abstract

STUDY DESIGN

An experimental study to investigate the activation of Src/Stat3 pathways in chordomas and blockage of this pathway as a potential strategy for chordoma treatment.

OBJECTIVE

To investigate the activation of Src/Stat3 pathway in chordomas cells and to determine the efficiency of inhibiting this pathway by 2-cyano-3,12-dioxooleana-1,9 (11)-dien-28-oic acid-methyl ester (CDDO-Me) as a potential chemotherapeutic agent for chordoma treatment.

SUMMARY OF BACKGROUND DATA

The advent of molecularly targeted therapies has raised interest for their use in the treatment of chordomas. Unfortunately, the current understanding of molecular markers in chordomas is limited. Constitutive activation of Stat3 is a common finding in a wide spectrum of human cancers. The function of Stat3 pathway in chordomas has not been elucidated.

METHODS

The expression of key components of the Src/Stat3 signaling cascade was evaluated by Western blot in chordoma tissues and chordoma cell lines. The effects of CDDO-Me on chordoma cell growth were evaluated in these chordoma cell lines by MTT assay. The expression of key components of the Src/Stat3 signaling cascade and poly (ADP-ribose) polymerase cleavage in these CDDO-Me treated cells were analyzed by Western blot and immunofluorescence. Furthermore, the synergistic effect of CDDO-Me on cisplatin and doxorubicin-induced cytotoxicity was evaluated by MTT. Finally, chordoma cells were grown in a 3-dimensional (3D) culture and treated with CDDO-Me.

RESULTS

The key components of the Src/Stat3 signaling cascade, including Stat3, pStat3, Src, pSrc, Bcl-xL, and Myeloid Cell Leukemia-1, were all highly expressed in chordomas. Expression of the key components of the Src/Stat3 signaling cascade was inhibited in chordoma cells after treatment with CDDO-Me. The growth of chordoma cells was inhibited and apoptosis associated poly (ADP-ribose) polymerase cleavage was detected after treatment with CDDO-Me. Additionally, CDDO-Me has a synergistic effect on cisplatin or doxorubicin-induced chordoma cell death (P < 0.001). Finally, expression of pSrc and pStat3 and chordoma cell growth was inhibited by treatment of CDDO-Me using 3D culture.

CONCLUSION

The Src/Stat3 signaling pathway was activated in chordomas. Blockage of Src/Stat3 pathway by CDDO-Me is a potential strategy for chordoma treatment and may be focus for future research.

摘要

研究设计

一项实验研究，旨在探讨 Src/Stat3 通路在脊索瘤中的激活情况，并探讨阻断该通路作为脊索瘤治疗的潜在策略。

目的

探讨 Src/Stat3 通路在脊索瘤细胞中的激活情况，并确定 2-氰基-3,12-二氧代-1,9(11)-二烯-28-酸甲酯（CDDO-Me）作为潜在化疗药物抑制该通路的效率。

背景资料概要

分子靶向治疗的出现引起了人们对其在脊索瘤治疗中的应用的兴趣。不幸的是，目前对脊索瘤分子标志物的了解有限。Stat3 的组成性激活是广泛存在于人类癌症中的一种常见现象。Stat3 通路在脊索瘤中的功能尚未阐明。

方法

通过 Western blot 评估脊索瘤组织和脊索瘤细胞系中 Src/Stat3 信号级联的关键成分表达。通过 MTT 测定评估 CDDO-Me 对这些脊索瘤细胞系中脊索瘤细胞生长的影响。通过 Western blot 和免疫荧光分析这些 CDDO-Me 处理细胞中 Src/Stat3 信号级联的关键成分和多聚（ADP-核糖）聚合酶切割的表达。此外，通过 MTT 评估 CDDO-Me 对顺铂和阿霉素诱导的细胞毒性的协同作用。最后，将脊索瘤细胞在 3 维（3D）培养中生长并进行 CDDO-Me 处理。

结果

Src/Stat3 信号级联的关键成分，包括 Stat3、pStat3、Src、pSrc、Bcl-xL 和髓样细胞白血病-1，在脊索瘤中均高度表达。CDDO-Me 处理后，脊索瘤细胞中 Src/Stat3 信号级联的关键成分表达受到抑制。CDDO-Me 处理后，脊索瘤细胞生长受到抑制，凋亡相关多聚（ADP-核糖）聚合酶切割被检测到。此外，CDDO-Me 对顺铂或阿霉素诱导的脊索瘤细胞死亡具有协同作用（P<0.001）。最后，通过 3D 培养用 CDDO-Me 处理，抑制 pSrc 和 pStat3 的表达和脊索瘤细胞生长。